Purification and some properties of ornithine decarboxylase from rat liver.
Ornithine decarboxylase (EC 4.1.1.17) was purified to near homogeneity from the livers of thioacetamide- and DL-alpha-hydrazino-delta aminovaleric acid-treated rats by using three types of affinity chromatography with pyridoxamine phosphate-Sepharose, pyridoxamine phosphate-dipropylenetriamine-Sepharose and heparin-Sepharose. This procedure gave a purification of about 3.5.10(5)-fold with an 8% yield; the specific activity of the final enzyme preparation was 1.1.10(6) nmol CO2/h per mg protein. The purified enzyme gave a single band of protein which coincided with activity peak on polyacrylamide gel electrophoresis and also gave a single major band on SDS-polyacrylamide gel electrophoresis. A single precipitin line was formed between the purified enzyme and an antiserum raised against a partially purified enzyme, on Ouchterlony immunodiffusion. The molecular weight of the enzyme was estimated to be 105000 by polyacrylamide gel electrophoresis at several different gel concentrations; the dissociated subunits had molecular weights of 50000 on SDS-polyacrylamide gels. The isoelectric point of the enzyme was pH 4.1.
D000818:Animals; D002262:Carboxy-Lyases / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D004790:Enzyme Induction; D005779:Immunodiffusion; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000201:enzymology*; D008297:Male; D009952:Ornithine / Q000031:analogs & derivatives / Q000494:pharmacology; D009955:Ornithine Decarboxylase / Q000302:isolation & purification* / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013853:Thioacetamide / Q000494:pharmacology
D013853:Thioacetamide; C007970:alpha-hydrazinoornithine; D009952:Ornithine; D002262:Carboxy-Lyases; D009955:Ornithine Decarboxylase